P1649: STUDIES OF COMPLEX PRODUCTS BASED ON HEPARINS OF ANIMAL AND PLANT ORIGIN
Authors: Margarita Lyapina, Marina Kalugina, Evgenia Maystrenko
Journal: HemaSphere (2023)

Topic: 34. Thrombosis and vascular biology - Biology & Translational Research Background: It is known that many plants serve as a source of medicinal raw materials for the production of anticoagulants, fibrinolytics and thrombolytics. The development of preparations of low-molecular-weight heparins (LMH) of different nature without adverse side effects on the human body is currently becoming particularly relevant. The presence of fractions of low molecular weight heparin in the roots of tree-like peonies has been established. Aims: The purpose of this work is to create complex products including low molecular weight heparins (LMH) of animal and vegetable origin together with an amino acid (glutamic acid– GA), to study their effect on blood clotting and to identify the most effective anti-clotting properties of the compound LMH-GA. Methods: Peony roots served as raw materials for the production of plant heparin-like substances. Extracts from P.lutea were used as herbal preparations, and the standard commercial drug Fraxiparine (France) was used as a comparison drug. The presence of heparin-like components was determined using chemical and photoelectrocolorimetric methods. All animal experiments were carried out in accordance with ethical principles and documents recommended by the Helsinki Declaration on Humane Treatment of Animals. For the research, blood was taken from the jugular vein (vena jugularis) using 3.8% sodium citrate as a preservative in a ratio of 9:1. To obtain platelet-poor plasma, blood was centrifuged at 3000g for 10-12 minutes. The anticoagulant activity of plasma was judged by the increase in clotting time in the tests of activated partial thromboplastin time (APTT), which characterizes the internal mechanism of blood clotting, thrombin time, reflecting the general path of blood clotting. To characterize the parameters of the fibrinolytic link of the anticoagulation system of blood in the blood plasma, the total fibrinolytic activity was determined, including the activity of heparin complexes with blood plasma components and plasmin activity; fibrin depolymerization activity (PDPA), reflecting the processes of fibrin depolymerization. Complex heparin agents of animal and vegetable origin with the inclusion of glutamic acid in their composition were obtained. Solutions of each of the heparins were combined with a solution of glutamic acid in a molar ratio of 1:4 and their effect on the state of plasma hemostasis was studied in vitro. Results: It is shown in all compounds (plant heparinoid and standard LMH) the presence of anticoagulant properties, which is realized by an increase in APTT by 2.2 times (heparinoid) and by 1.9 times (Fraxiparine) in comparison with the control (NaCl). This is due to their anti-factor Xa activity. Connections heparinoid with GA, the APTT was increased by 3.1 times, while Fraxiparine with GA showed weaker anticoagulant activity, increasing APTT only by 2.4 times. Thrombin time in all compounds did not change. It has also been shown that all compounds of heparins with glutamic acid interfere with the processes of fibrin formation due to the presence of fibrin-depolymerization activity in them, and the compound of heparinoid with GA had a more significant PDPA, exceeding the similar effect of the compound Fraxiparine with GA by 23-25%. Summary/Conclusion: For the first time, the ability of low-molecular-weight heparin from peony roots together with glutamic acid to exhibit synergistic anticoagulant and fibrin-depolymerization effects exceeding those of heparin alone in the same concentrations has been established. The predominant anticoagulant effect of a plant-derived heparinoid compound is shown, rather than a compound of standard heparin of animal origin. The results obtained may undoubtedly be of interest. Keywords: Fibrinolysis, Heparin, Thrombosis, Antithrombotic